Abstract
Endo-β-N-acetylglucosaminidase D (Endo D) produced by Streptococcus pneumoniae hydrolyzes the di-N-acetylchitobiose structure in the core of complex-type asparagine-linked oligosaccharides, and has a molecular weight of 180 kDa. A truncated Endo D of 102 kDa in which 134 N-terminal amino acids and 599 C-terminal amino acids were deleted, still retained the enzymatic activity. The truncated Endo D has specificity indistinguishable from the intact enzyme, and also acted on the core structure of asparagine-linked oligosaccharides attached to intact IgG. Because of its lower molecular weight, the truncated enzyme may be useful as a tool for protein deglycosylation. The entire region of the truncated Endo D had 32% sequence identity to endo- β-N-acetylglucosaminidase BH (Endo BH) from Bacillus halodurans, which acted on high-mannose type oligosaccharides. Chimeric constructs of the truncated Endo D and Endo BH showed no activity. Glutamic acid 324 (E 324) in Endo D is conserved in Endo BH and Endo M, and is an essential amino acid in Endo M. Mutation of E324 abolished Endo D activity. The specificity of Endo D for complex type oligosaccharides is probably defined by multiple domains in the Endo D structure. Published in 2005.
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Abbreviations
- DE:
-
delayed extraction
- Endo:
-
endo-β-N-acetylglucosaminidase
- LC:
-
liquid chromatography
- MS:
-
mass spectrometry
- MALDI-TOF:
-
matrix-assisted laser desorption ionization-time of flight
- PBS:
-
Dulbecco’s phosphate buffered saline
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Yamamoto, S., Muramatsu, H. & Muramatsu, T. Mutational studies on endo-β-N-acetylglucosaminidase D which hydrolyzes core portion of asparagine-linked complex type oligosaccharides. Glycoconj J 22, 35–42 (2005). https://doi.org/10.1007/s10719-005-0847-7
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DOI: https://doi.org/10.1007/s10719-005-0847-7